Involvement of Abnormal p-α-syn Accumulation and TLR2-Mediated Inflammation of Schwann Cells in Enteric Autonomic Nerve Dysfunction of Parkinson's Disease: an Animal Model Study.

The study was designed to investigate the pathogenesis of gastrointestinal (GI) impairment in Parkinson's disease (PD). We utilized 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP, 20 mg/kg) and probenecid (250 mg/kg) to prepare a PD mice model. MPTP modeling was first confirmed. GI motility was measured using stool collection test and enteric plexus loss was also detected. Intestinal phosphorylated α-synuclein (p-α-syn), inflammation, and S100 were assessed using western blotting. Association between Toll-like receptor 2(TLR2) and GI function was validated by Pearson's correlations. Immunofluorescence was applied to show co-localizations of intestinal p-α-syn, inflammation, and Schwann cells (SCs). CU-CPT22 (3 mg/kg, a TLR1/TLR2 inhibitor) was adopted then. Success in modeling, damaged GI neuron and function, and activated intestinal p-α-syn, inflammation, and SCs responses were observed in MPTP group, with TLR2 related to GI damage. Increased p-α-syn and inflammatory factors were shown in SCs of myenteron for MPTP mice. Recovered fecal water content and depression of inflammation, p-α-syn deposition, and SCs activity were noticed after TLR2 suppression. The study investigates a novel mechanism of PD GI autonomic dysfunction, demonstrating that p-α-syn accumulation and TLR2 signaling of SCs were involved in disrupted gut homeostasis and treatments targeting TLR2-mediated pathway might be a possible therapy for PD.

Phosphorylated α-synuclein aggregated in Schwann cells exacerbates peripheral neuroinflammation and nerve dysfunction in Parkinson's disease through TLR2/NF-κB pathway.

To investigate the mechanism of peripheral neuropathy in Parkinson's disease (PD), we prepared a PD mice model by long-term exposure of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to mimic PD pathology in humans and the sciatic nerves were taken for further research. It turned out that phosphorylated α-synuclein (p-α-syn) was significantly deposited in Schwann cells (SCs) of sciatic nerves possibly contributing to degenerated myelin SCs and atrophied axons in MPTP group. Further analysis confirmed that toll-like receptors (TLRs) were implicated with PD peripheral neuropathy, in which TLR2 exhibits the predominant expression. Increased expression of inflammatory factors about TLR2/nuclear factor kappa-B (NF-κB) pathway was noted in MPTP group compared to saline group, with proteins on other pathways showing no changes. Moreover, MPTP-challenged mice exhibited worse motor ability and damaged nerve conduction, implicating that p-α-syn neurotoxicity might be relevant to impairments of motor and sensory nerves. After the treatment of CU-CPT22, a TLR2 antagonist, p-α-syn accumulation, motor and sensory function were ameliorated in CU-CPT22 combined with MPTP group. Thus, we demonstrated that pathological p-α-syn might combine TLR2 to affect SCs activation, inflammatory response as well as motor and sensory function through TLR2/nuclear factor kappa-B (NF-κB) signaling pathway. This study firstly demonstrates a novel mechanism of p-α-syn accumulated in SCs of peripheral nerves, which extends our understanding on SCs-mediated peripheral neuroinflammation related to TLR2/NF-κB signaling pathway and sheds light on potential new therapeutic avenues for PD.

Phosphorylated α-synuclein and phosphorylated tau-protein in sural nerves may contribute to differentiate Parkinson's disease from multiple system atrophy and progressive supranuclear paralysis.

Differential diagnosis of Parkinson's disease (PD), multiple system atrophy (MSA) and progressive supranuclear paralysis (PSP) is challenging. This study aimed to investigate the expression of phosphorylated α-synuclein (p-α-syn) and phosphorylated tau-protein (p-tau) in sural nerves from patients with PD, MSA and PSP to find biomarkers for differential diagnosis. Clinical evaluations and sural nerve biopsies were performed on 8 PD patients, 8 MSA patients, 6 PSP patients and 8 controls (CTRs). Toluidine blue staining was used to observe morphological changes in sural nerves. The deposition of p-α-syn and p-tau was detected by immunohistochemistry with semiquantitative evaluation. Locations of p-α-syn and p-tau were identified by double immunofluorescent staining. In case groups, the density of nerve fibres decreased with swollen or fragmented Schwann cells (SCs). All cases (22/22) but no CTRs (0/8) presented p-α-syn immunoreactivity with gradually decreasing semiquantitative levels among the PD (6.00 ± 2.07), MSA (5.00 ± 2.33) and PSP (3.50 ± 1.52) groups. p-tau aggregates were found in 7/8 MSA (1.88 ± 1.46) and 6/6 PSP (1.67 ± 0.52) patients but not in PD patients or CTRs. There were different expression patterns of p-α-syn and p-tau in PD, MSA and PSP patients. These findings suggest that peripheral sensory nerve injury exists in PD, MSA and PSP patients. With a different expression pattern and level, p-α-syn and p-tau in sural nerves may serve as novel biomarkers for differential diagnosis of PD, MSA and PSP.

Clusterin is associated with spontaneous breast cancer in TA2 mice.

Two-dimensional electrophoresis and Matrix-assisted laser desorption ionization-time of flight-time of mass spectrometry were used to detect the differentially expressed proteins in serum of tientsin albinao 2 mice with spontaneous breast cancer, normal tientsin albinao 2 mice and tientsin albinao 1 mice. Only nuclear clusterin (n-CLU) was expressed in tientsin albinao 1. Immunohistochemistry and western blot validated that n-CLU was present in normal tientsin albinao 2 and tientsin albinao 1 mammary epithelium, and secretory clusterin expressed in the cytoplasm of normal tientsin albinao 2 mammary epithelium and spontaneous breast cancer. n-CLU may play an important role in tientsin albinao 2 spontaneous breast cancer initiation and development.

[The influence of different microenvironments on melanoma invasiveness and microcirculation patterns].

OBJECTIVE: To investigate the influence of different microenvironments on tumor microcirculation patterns and invasive capability. METHODS: Melanoma B16 cells were injected into the peritoneal cavity and skeletal muscle of C57 mice synchronously. CK18 expression in melanoma was assessed to distinguish the malignant phenotype of tumors in the peritoneal cavity from that in the skeletal muscle. HIF-1alpha, MMP-2 and MMP-9 protein and mRNA expression were compared in the two microenvironments. Cells positive for each immunohistochemical stain and the vessels representative of each type of microcirculation pattern were evaluated in two microenvironments. RESULTS: CK18 and HIF-1alpha expression in melanoma were significantly higher in the skeletal muscle than in the peritoneal cavity (t = 8.142, t = 3.645, P < 0.05). Compared with the peritoneal cavity, melanoma cells in the skeletal muscle overexpressed MMP-2 and MMP-9 (t = 4.916, t = 7.782, P < 0.05). Real time-PCR results also showed that MMP-2 and MMP-9 mRNA levels in melanoma were higher in the skeletal muscle than in the peritoneal cavity (t = 36.814, t = 26.025, P < 0.05). Vasculogenic mimicry channels and endothelium-dependent vessels were the major microcirculation patterns in the skeletal muscle and in the peritoneal cavity respectively. CONCLUSIONS: Different microenvironments affect invasiveness and blood supply patterns of melanoma. Different microenvironment induced tumor cell secretion of more invasion-related proteins and affect invasiveness and blood supply patterns of melanoma.

[The effect of 20(R)-ginsenoside Rg3 on the differential expression of cell signaling genes and other related genes in human lung adenocarcinoma cell line A549].

OBJECTIVE: To study the effect of 20(R)-ginsenoside Rg3 [20(R)-Rg3] on the differential expression of cell signaling genes and other related genes in human lung adenocarcinoma cell line A549. METHODS: The cell line A549 was cultured with 20(R)-Rg3 (10(-6) mol/L) for 72 h, RNA was extracted, and the differential expression of cell signaling genes and other related genes were detected with DNA microarray. RESULTS: A total of 3 490 genes were detected, and the expression of 24 genes were changed after the addition of 20(R)-Rg3. Two cell signaling genes & transducin genes were up-regulated. Two cystoskeleton & locomotion genes were up-regulated. Three proto-oncogene & anti-oncogene genes were down-regulated. Two of the translation & synthetical genes were down-regulated, and 1 of them was up-regulated. One DNA recombination gene and one metabolism gene were down-regulated. CONCLUSION: 20(R)-Rg3 showed significant effect on the differential expression of cell signaling genes and other related genes in human lung cell line A549.

[Research on the laser atomization treatment machine].

This text has introduces a new-type laser treatment device. It utilizes the ultrasound atomized gas passage as its optics and makes the laser beams together with the atomized medicine to be transmitted to the patient's respiratory track and lungs for treatment.

Combined treatment of glibenclamide and CoCl2 decreases MMP9 expression and inhibits growth in highly metastatic breast cancer.

BACKGROUND: To observe the influence of combination treatment with glibenclamide and CoCl(2) on the growth and invasiveness of TA2 breast cancer, and to detect the protein and mRNA expression of MMP9. METHODS: 50 adult female TA2 mice were randomly divided into 5 groups including DMSO control, CoCl(2), glibenclamide, CoCl(2) + glibenclamide and paclitaxel. All of these mice were inoculated with TA2 spontaneous breast cancer cells in the left groin. Nine days after inoculation the tumor could be palpated. Different treatments for each group were then subcutaneously administered near the tumors on the 9th and 14th days after injection. Tumor size was measured to determine the growth curve. All mice were sacrificed on the 18th day after initial inoculation and tumor tissues were collected. Some fresh tissues without necrosis were stored at -80°C for mRNA detection and the other tumor tissue was fixed with 10% formalin for H&E and immunohistochemical staining. RESULTS: The growth rate of tumor cells in the CoCl(2) + glibenclamide group was lower than that seen in the other groups. On the 14th day, the average volume of tumor in the CoCl(2) + glibenclamide group was the lowest and the difference has statistical significance (P < 0.05), while the differences among the CoCl(2), glibenclamide and paclitaxel had no statistical significance. The mean percentage of cells expressing MMP9 and PCNA was the lowest in the CoCl(2) + glibenclamide group (P < 0.05). MMP9 mRNA expression paralleled MMP9 protein expression in these groups (P < 0.05). CONCLUSIONS: Combined treatment with glibenclamide and CoCl(2) inhibits TA2 spontaneous breast cancer growth and invasiveness with effects similar to paclitaxel.

Number of glioma polyploid giant cancer cells (PGCCs) associated with vasculogenic mimicry formation and tumor grade in human glioma.

BACKGROUND: Polyploid giant cancer cells (PGCCs) contribute to solid tumor heterogeneity. This study investigated the relationships among PGCCs numbers, vasculogenic mimicry (VM) formation, and tumor grades in glioma. METHODS: A total of 76 paraffin-embedded glioma tissue samples, including 28 cases of low grade and 48 cases of high grade gliomas, were performed with H&E and immunohistochemical staining for Ki-67 and hemoglobin. The size of PGCCs nuclei was measured by a micrometer using H&E section and defined as at least three times larger than the nuclei of regular diploid cancer cells. The number of PGCCs and different blood supply patterns were compared in different grade gliomas. Microcirculation patterns in tumors were assessed using CD31 immunohistochemical and PAS histochemical double staining. Human glioma cancer cell line C6 was injected into the chicken embryonating eggs to form xenografts, which was used to observe the PGCCs and microcirculation patterns. RESULTS: In human glioma, the number of PGCCs increased with the grade of tumors (χ2 = 4.781, P = 0.015). There were three kinds of microcirculation pattern in human glioma including VM, mosaic vessel (MV) and endothelium dependent vessel. PGCCs were able to generate erythrocytes via budding to form VM. The walls of VM were positive (or negative) for PAS staining and negative for CD31 staining. There were more VM and MVs in high grade gliomas than those in low grade gliomas. The differences have statistical significances for VM (t = 3.745, P = 0.000) and MVs (t = 4.789, P = 0.000). PGCCs, VM and MVs can also be observed in C6 chicken embryonating eggs xenografts. CONCLUSIONS: The data demonstrated presence of PGCCs, VM and MVs in glioma and PGCCs generating erythrocytes contribute the formation of VM and MVs.